This antibody can be used for WB, but the signal is very faint, and we do not recommend or guarantee it for this method. We recommend the clone MEM-255 (ab9527) or the rabbit polyclonal ab14989, which are much better for WB.

Post-translationalmodificationsPalmitoylated.Phosphorylated by FYN on Tyr-317 in resting T-cells; which promotes interaction with CSK. Dephosphorylated by PTPRC/CD45 upon TCR activation; which leads to CSK dissociation. May also be dephosphorylated by PTPN11. Hyperphosphorylated in mast cells upon FCER1 activation.

Anti-PAG antibody [PAG-C1] images

Flow Cytometry - Anti-PAG antibody [PAG-C1] (ab4206)

Overlay histogram showing Jurkat cells stained with ab4206 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab4206, 1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H+L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

The immunoprecipitation used covalently bound antibody to Sepharose beads as immunosorbent. This method of precipitation obviously differs from proteinA/G-sepharose or GoatxMs -Sepharose mediated immunoprecipitation protocols.